20-hydroxyecdysone accumulation and regulation in Ajuga lobata D. Don suspension culture

Figures & data

Fig. 1. Cell dry weight in A. lobata D. Don culture (Spots indicate the cell dry weight at different culture times, curve is cell dry weight kinetics).

Fig. 2. Dynamic changes in cell suspension electric conductivity and 20E content.

Notes: 2-a 20E content in A. lobata D. Don culture (Spots indicate the 20E production at different culture times, curve is 20E accumulation kinetics); 2-b Dynamic changes of electric conductivity in cell suspension.

Fig. 3. Dynamic change of nutrients in A. lobata D. Don culture.

Notes: 3-a Sugar content kinetics of A. lobata D. Don culture; 3-b Phosphate salt change in A. lobata D. Don culture; 3-c Nitrate salt change in A. lobata D. Don culture; 3-d Ammonium salt change in A. lobata D. Don culture.

Fig. 4. Influence of passaging on 20E content in suspension culture.

Table 1. Effect of L-PAL on A. lobata D. Don cell suspension culture.

L-PAL concentration (mmol L^−1)	Cell growth	Cell activity MV ± SD	20E (mg g^−1) MV ± SD
0	+	0.32 ± 0.04	2.18 ± 0.08
0.1	+	0.58 ± 0.05**	2.61 ± 0.66
0.2	+	0.78 ± 0.09**	2.80 ± 0.29
0.4	+	0.66 ± 0.06**	2.71 ± 0.19

Note: +: Cell is growing and medium is gray-white in color. MV: mean values; SD: standard deviation.

^** p < 0.01 in comparison with control cultures.

Table 2. Effect of MVA on A. lobata D. Don cell suspension culture.

MVA concentration (mg·L^-1)	Cell growth	Cell activity MV ± SD	20E(mg·g^−1) MV ± SD
0	+	0.32 ± 0.04	2.18 ± 0.08
5	+	0.38 ± 0.03	5.25 ± 2.06*
10	+	0.38 ± 0.05**	11.75 ± 1.51**
20	–	0.43 ± 0.06**	4.25 ± 0.42
80	–	–	–

Notes: +: cell is growing and medium is gray-white in color; – cell growth is inhibited or cells are dead. MV: mean values; SD: standard deviation.

^* p < 0.05;

^** p < 0.01 in comparison with control cultures.

Table 3. Effect of α-Pinene on A. lobata cell suspension culture.

α-Pinene concentration (μl L^−1)	Cell growth	Cell activity MV ± SD	β-EC (mg g^-1) MV ± SD
0	+	0.32 ± 0.04	2.18 ± 0.08
25	+	0.28 ± 0.04	2.75 ± 0.52**
50	+	0.60 ± 0.06**	3.50 ± 0.14**
100	–	0.39 ± 0.05**	3.25 ± 0.12**

Notes: +: cell are growing and medium is gray-white in color; – cell growth is inhibited or cells are dead. MV: mean values; SD: standard deviation.

^** p < 0.01 in comparison with control cultures.

Table 4. Effect of terpineol on A. lobata cell suspension culture.

Terpineol concentration (μl L^−1)	Cell growth	Cell activity MV ± SD	β-EC (mg g^−1) MV ± SD
0	+	0.32 ± 0.04	2.18 ± 0.08
100	+	0.30 ± 0.06	5.33 ± 0.73**
400	–	–	1.795 ± 0.51
600	–	–	1.95 ± 0.40

Notes: +: cell are growing and medium is gray-white in color; – cell growth is inhibited or cells are dead. MV: mean values; SD: standard deviation.

^** p < 0.01 in comparison with control cultures.

Fig. 5. Effect of NO on 20E content.

Note: ** p < 0.01; * p < 0.05 in comparison with control cultures.

Table 5. Effect of different combination of exogenous substrates on A. lobata cells.

Substrate	Cell fresh weight( g) MV ± SD	20E (mg·g^-1) MV ± SD
CK	26.74 ± 1.87	2.18 ± 0.08
α-Pinene 50 μl L^-1	24.93 ± 0.32	3.50 ± 0.14*
SNP 80 μmol L^−1	27.98 ± 0.56	2.84 ± 0.75
MVA 10 mg L^−1	25.89 ± 0.31	11.75 ± 1.51**
MVA + SNP	27.11 ± 0.60	12.58 ± 0.12**
α-Pinene + SNP	27.78 ± 1.05	9.25 ± 1.21**
MVA + α-Pinene	25.15 ± 0.68	9.25 ± 1.35**
MVA + α-Pinene + SNP	25.44 ± 0.69	12.75 ± 0.75**

Notes: CK: Supplementation of passage 10 A. lobata D. Don cell culture; MV: mean values; SD: standard deviation.

^* p < 0.05;

^** p < 0.01 in comparison with control cultures.